宿主蛋白HDAC3调控VSV复制的机制研究

水疱性口炎病毒（VSV）具有广泛的细胞嗜性,主要原因之一是其可借助宿主蛋白参与对机体代谢系统的调控来保障病毒自身的复制周期。本课题组前期研究发现，抑制/沉默宿主细胞组蛋白去乙酰化酶3（HDAC3）及沉默与HDAC3互作的核受体共抑制因子1（NCOR1）均可降低VSV复制，提示HDAC3在染色质水平参与调控作用，但该分子如何影响VSV复制尚不清楚。鉴于此，本项目拟过表达与敲除Hdac3基因，验证其对VSV复制的影响；利用激光共聚焦及Western blot明确VSV感染对HDAC3定位及表达的影响；利用染色质免疫共沉淀-测序，解析VSV感染后HDAC3在全基因组DNA互作位点的变化，鉴定受调控的关键基因；利用免疫共沉淀及质谱技术，明确与HDAC3互作并协同调控转录的蛋白。该项目的预期结果不仅有利于阐明HDAC3对VSV复制增殖的调控机制，而且也为寻找潜在阻止VSV感染的靶点提供理论依据。

Vesicular stomatitis virus (VSV) has broad cell tropism, one of main reasons is that VSV replicate and propagate by harnessing host metabolism system via host conserved proteins. Based our pilot study, inhibiting/silencing Histone deacetylase 3 (HDAC3) and silencing Nuclear receptor co-repressor 1(NCOR1), which interacts with HDAC3 in the nucleus, could inhibit VSV replication, suggesting HDAC3 functions in chromosome level to regulate VSV replication. However, how HDAC3 regulates the replication of VSV is still largely elusive. For this reason, this project will test the influence of HDAC3 on VSV replication by overexpressing or knocking out Hdac3 gene; make sure whether or how VSV infection affects HDAC3 distribution and expression change by laser confocal fluorescence microscope and Western blot; study HDAC3 enrichment region change across the whole genome through Chromatin Immunoprecipitation sequencing technique, which is followed by identifying the key genes; study the key protein interacted cooperatively with HDAC3 to regulate gene transcription during VSV infection through Co-Immunoprecipitation and Mass Spectrometry technique. The foreseen results will not only uncover the mechanism how HDAC3 regulates VSV replication and propagation, but also provide theoretical guide to search for targets to block VSV infection.